Kumar Abhinendra, Mhatre Sharayu, Dikshit Rajesh
Centre for Cancer Epidemiology, Advanced Centre for Treatment Research and Education in Cancer, Tata Memorial Centre; Homi Bhabha National Institute (HBNI), Training School Complex, Anushaktinagar, Mumbai, Maharashtra, India.
Indian J Med Microbiol. 2019 Oct-Dec;37(4):514-520. doi: 10.4103/ijmm.IJMM_19_441.
Identifying infectious pathogens by collecting intravenous blood (IVB) is a well-established procedure, however, the collection of IVB in field epidemiological study is challenging. The dried blood spot (DBS) as an alternative to IVB has been introduced, although, there is a limited study to demonstrate the utility of DBS stored at various storage conditions and transported at different periods. This is an observational study, which evaluates the effectiveness of DBS in field epidemiological studies to identify infectious pathogens.
A total of 264 paired DBS samples prepared from IVB, stored at 4°C, -20°C after period 24, 48 and 72 h. Serologically, enzyme-linked immunosorbent assay [ELISA] IgG antibody detected against Helicobacter pylori infection from DBS and compared with IVB.
Quantitatively, IgG antibody reactivity showed >87% correlation between IVB and DBS samples stored at 4°C or -20°C within 48 h of transport duration. DBS stored at 4°C shows, equal sensitivity 87.5% and specificity 95% before 48 h of transport duration, while at -20°C storage similar sensitivity 87.5% observed but slightly less specificity 86.36% observed as compared to 24 h of transport duration. One-way analysis of variance showed, nonsignificant difference at both (-20°C and 4°C) the stored condition with P value (P > 0.851) and (P > 0.477). Kappa values showed good inter-rater reliability between DBS and IVB in a range (0.77-0.81).
No significant difference was observed in detecting H. pylori when ELISA was conducted using IVB or DBS stored at 4°C and transported even after 48 h. This confirms that DBS collected even in compromised conditions in the field can be used for detecting infection.
通过采集静脉血来鉴定感染性病原体是一种成熟的方法,然而,在现场流行病学研究中采集静脉血具有挑战性。已引入干血斑(DBS)作为静脉血的替代方法,不过,关于在各种储存条件下储存并在不同时间段运输的DBS的效用的研究有限。这是一项观察性研究,评估DBS在现场流行病学研究中鉴定感染性病原体的有效性。
总共264对由静脉血制备的DBS样本,在24、48和72小时后分别储存在4°C、-20°C。通过血清学方法,采用酶联免疫吸附测定(ELISA)检测DBS中针对幽门螺杆菌感染的IgG抗体,并与静脉血进行比较。
定量分析显示,在运输时间48小时内,储存在4°C或-20°C的静脉血和DBS样本之间IgG抗体反应性的相关性>87%。储存在4°C的DBS在运输时间48小时之前显示出相同的敏感性87.5%和特异性95%,而在-20°C储存时观察到相似的敏感性87.5%,但与运输24小时相比,特异性略低,为86.36%。单因素方差分析显示,在两种储存条件(-20°C和4°C)下差异均无统计学意义,P值分别为(P>0.851)和(P>0.477)。kappa值显示DBS和静脉血之间的评分者间可靠性良好,范围为(0.77-0.81)。
使用静脉血或储存在4°C甚至运输48小时后的DBS进行ELISA检测时,在检测幽门螺杆菌方面未观察到显著差异。这证实即使在现场条件不佳的情况下采集的DBS也可用于检测感染。